Long-term objective: to formulate a general model for the pathogenesis of hepatic fibrosis at the molecular level. Better understanding of changes in protein synthesis in cirrhotic liver will permit development of diagnostic methods to evaluate fibrogenesis in man. Specific aims: (1) To determine changes in hepatic protein synthesis caused by chronic ethanol administration to baboons which have the entire spectrum of alcoholic liver disease. (2) To evaluate the level of gene regulation responsible for differences in collagen mRNA in cirrhotic liver as compared to control liver. (3) To identify which cell(s) is primarily responsible for Type I collagen synthesis in alcoholic liver disease in baboons. (4) To investigate the influence of fibronectin synthesis as an initiator of fibrosis in liver. (5) To evaluate collagen synthesis by a noninvasive serum assay and correlate Type I collagen carboxy-terminal propeptide RIA with collagen mRNA sequence content infibrotic human liver biopsies. (6) To evaluate the possibility that a species' propensity for developing cirrhosis may result from a collagen gene polymorphism. (7) To determine whether any collagen gene polymorphism exists in man which may be related to development of cirrhosis in only a fraction of chronic ethanol users. Methods: cell-free protein synthesis, immunoprecipitation of protein products, "Northern" transfer and "Dot" blot hybridization will be employed to study the effect of chronic ethanol administration on RNA extracted from liver biopsy specimens in baboons. Wedge biopsy specimens from cirrhotic and control baboons will be studied by standard in vitro transcription assays to determine whether transcriptional regulation is responsible for changes in collagen mRNAs in fibrotic liver. In situ hybridization using a Type I procollagen cDNA probe, isolated hepatocytes and tissue sections from fibrotic and control liver will be used to localize the cell(s) which is primarily responsible for collagen synthesis. Investigation of collagen gene polymorphism will involve structural evaluation of Type I collagen genes using Southern blot hybridization and DNA sequencing of genomic DNA extracted from white blood cells of human and animal ethanol abusers. Health relatedness: determining a positive correlation between the collagen propeptide RIA and procollagen mRNA sequence content will help validate RIA as a non-invasive method for assessing prognosis in chronic liver disease. Identification of human collagen gene polymorphism may be useful in predicting which alcohol abusers are at a high risk to develop cirrhosis.